Malignant tumors are known to have many different types of cells in it. These cells have genes and proteins that are very different from one another. And they grow at different rates. This is known as heterogeneity. The heterogeneity is also responsible for combining chemotherapy with radiotherapy and/or various kind of chemotherapy in combination for effective treatment of malignant tumors.
There is a lack of well defined antigens for organ/tissue specific cancer. To overcome this problem cancer cells are used as an antigen. The use of cancer cells provides benefit of repertoires of the antigens present on cancer cells.
The cancer cells can be sourced from the same patients (autologous) or from a different patient (allogeneic).
Use of autologous cancer cell in vaccines is personalized therapy and is associated with practical difficulties. The autologous cells may not be available in all patients. When available it may not be of the desired quality and/or quantity. The approach is also time consuming. The approach is also associated with regulatory hurdles.
Use of allogeneic cancer cells is attractive as an antigen in therapeutic vaccine. However it suffers from lack of common antigen/s as cancer cells from a tissue/organ are heterogeneous in nature. The allogeneic cancer cells fail to elicit immune response against heterogeneous cancer cells specific to a tissue/organ. e.g. Allogeneic cell lines of pancreatic cancer Mia-paca-2 and Panc-1 produce immune response against themselves. However Mia-pica-2 cell line fails to elicit immune response against Panc-1 and Panc-1 fails to elicit immune response against Mia-paca-2
This can be overcome by use of multiple heterogeneous allogeneic cancer cells in a vaccine or identifying antigen present in a cancer tissue and using specific vaccine against it.
The heterogeneity of tumor makes it difficult to have a therapeutic vaccine with a single antigen to provide immune response against all the cells/majority of cells contained in the tumor. For this reason one need to combine allogeneic cells/antigens for therapeutic vaccine to make it effective against the tumor as a whole.
To overcome the problem of heterogeneity of the cancer/tumors, it is demonstrated to use more than one cell as antigen. Emens et al demonstrated use of more then one allogeneic cell line to cover the antigen repertoire of the heterogenic tumor/cancer. (Emens L A et al; J Clin Oncol. 2009 Dec. 10; 27(35):5911-8). While Laheru D et al demonstrated use of GM CSF to improve immunogenicity of the allogeneic cancer cells vaccine for treatment of cancer. Clin Cancer Res. 2008 Mar. 1; 14(5):1455-63).
Formalin-fixed tumor cells effectively induce anti-tumor immunity both in prophylactic and therapeutic conditions was explained by Chikage Obata, in Journal of dermatological science, Volume 34, issue 2, Pages 209-219 (May 2004) while a Clinical trial of autologous formalin-fixed tumor vaccine for glioblastoma multiform patients studied by Ishikawa E, in Cancer Sci. 2007 August; 98(8):1226-33, Epub 2007 May 22. In both the studies the efficacy is against the homologous cancer cells/tumors but none have demonstrated the killing of hetrogenous cancer cells specific to tissue/organ are killed by the vaccine.
Thus there is a need to have therapeutic vaccine using allogeneic cells as antigen for use in treatment of cancers which elicits immune response against heterogeneous cancer antigen/s specific to tissue/organ. E.g. therapeutic vaccine for pancreatic cancer using Mia-paca-2 cell line elicits immune response against Panc-1 and other pancreatic cancer cells.
Heterogeneous cancer cells specific to tissue/organ are those cancer cells which are present/originate from the same tissue/organ but fail to elicit and/or react to immune response generated by cancer cells which are present/originate from the same tissue/organ.
The methods for harvesting cancer cells and preserving them or propagating them are well known. The methods can be used for autologous as well as allogeneic cells, Some of the allogeneic cancer cell lines which are available for various type of tumors are listed below. The cell lines can be procured from various repositories like American Type Culture Collection, USA; Cell bank Australia, Australia; Coriell Cell Repositories, New Jersey USA; European Collection of Cell Cultures (ECACC), UK; German Collection of Microorganisms and Cell Cultures, Germany; Japanese Collection of Research Bioresources (JCRB), Japan; German Collection of Microorganisms and Cell Cultures, Germany; Korean Cell bank, Korea; RIKEN Bioresource Centre, Japan; Human Genetics Resource Center, USA; National Centre for Cell Science, India; MMRRC: Mutant Mouse Regional Resource Centers, USA; National Human Neural Stem Cell Resource, USA; UK Stem Cell Bank, UK and NCCS in India.
Also these or new cell lines or specific cancer cells can be isolated as described by Eton O, et al. Active immunotherapy with B irradiated Autologous whole Melanoma cells plus DETOX in patients with metastatic melanoma. In clinical cancer research, March 1998, Vol. 4, 619-627. Fresh tumor was collected at the time of surgery from frozen section laboratory and fragmented by slicing. to maximize the yield of viable tumor cells for vaccine preparation, the bulk of tumor was dissociated using collagenase type 1 (2 mg/ml) and type IV DNase (0.4 mg/ml) Sigma chemical Co., St Loius, Mo.; ref 25. These enzymes can alter the immunogenicity of the resulting cell preparation. The dissociated cells were washed in HBSS and gentamycin and resuspended in equal volumes of HBSS and chilled 10% DMSO+4% human serum albumin. Aliquots containing 1.5-2×10^7 viable tumor cells stored under liquid Nitrogen.
Robert O et al. described Irradiated Cells from Autologous Tumor Cell Lines as Patient-Specific Vaccine Therapy in 125 Patients with Metastatic Cancer: Induction of Delayed-Type Hypersensitivity to Autologous Tumor is Associated with Improved Survival in Cancer biotherapy and Radiopharmaceuticals Volume 17, Number 1, 2002. They established short-term cultures of pure tumor cells for use as autologous tumor cell vaccines in an effort to study the effects of patient-specific immunotherapy. Surgically resected fresh tumor was obtained from patients with metastatic cancer. Successful tumor cell lines (5×107) were expanded to 108 cells, irradiated, and cryopreserved for clinical use. Following a baseline test of delayed-type hypersensitivity (DTH) to an i.d. injection of 106 irradiated autologous tumor cells, patients received 3 weekly s.c. injections of 107 cells, had a repeat DTH test at week-4, then received monthly vaccinations for 5 months. A positive DTH test was defined as ≥10 mm induration; survival was determined from the first DTH test.
Dillman R O et al described Establishing in vitro cultures of autologous tumor cells for use in active specific immunotherapy in emphasis Tumor Immunol. 1993 July; 14(1): 65-9F They harvested fresh tumors and attempted to establish short-term cultures of tumor cells to obtain 10(8) cells which could subsequently be used in autologous tumor cell vaccine programs. Fresh tumors were mechanically processed to initiate primary cultures in RPM1-1640 containing 1 mM sodium pyruvate, 2 mM glutamine, 10 mM N-(2-hydroxyethyl) piperazine-N′-(2-ethanesulfonic acid), 15% fetal bovine serum, and antibiotics, incubated at 37 degrees C. in 5% CO2. We were successful in growing 87 of 142 [61%, (95% confidence limits [55-68%]) of all tumors] including 39 of 58 (67%) melanomas, 10 of 10 (100%) renal cell carcinomas, 14 of 14 (100%) sarcomas, and 23 of 54 (43%) various adenocarcinomas.
Jaffee E M described Development and characterization of a cytokine secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials in Cancer journal from scientific American, Vol. 4, issue 3, PP: 194. Freshly digested tumor cells were plated in duplicate at 2*10^6 cells per 25 cm2 flasks. Each growth condition was evaluated both separately and in combination with other growth supplements. Different media including RPMI, DMEM, Ham's and Aim V preparation, and lots of FBS were the initial components of growth media screened. After identification of the optimal medium and serum, additional additives were systematically evaluated. Each supplement was evaluated until either epithelial or fibroblastic like cells predominated in the cultures.
The book “Culture of animal cells—A manual of Basic technique”, Fifth edition, Protocol-24.3, pp: 429-430 also describes the methods of growing primary cells and tumors and establishing them as cell lines.
List of Cancer Cells Available from Various Repositories.
Cervical cancer: HeLa S3, HeLa 229, H1HeLa, Hs 588.T, GH329, GH354, HeLa NR1, C-4 I, C-4 II, DoTc2 4510, C-33 A, SW756 SiHa
Colon cancer: NCI-H548, Hs 255.T, HCT-8 (HRT-18), Hs 675.T
Bladder cancer: Hs 195.T, Hs 228.T, Hs 172.T5637, HT-1376 HT-1197, UM-UC-3, SW 780, J82 SCaBER, T24, TCCSUP, Hs 789.T, Hs 769.T, RT4
Renal Cancer: A704, A-704, NCI-H1373, NCI-H1395, Hs 618.T, SK-LU-1, HCC2935, HCC4006, HCC827, ACHN 786-O769-P, Caki-2 HTB-47, A-498 A549, A-427, SW 156, G-402, Hs 926.1, G-401
Breast Cancer: Hs 274.T, Hs 280.T, Hs 281.T, Hs 343.T, Hs 362.T, Hs 739.T, Hs 741.T, Hs 742.T, Hs 190.T Hs 319.T Hs 329.T Hs 344.T Hs 350.T Hs 371.T Hs 748.T Hs 841.T Hs 849.T Hs 851.T Hs 861.T Hs 905.T Hs 479.T, Hs 540.T, Hs 566(B).T, Hs 605.T, Hs 606 BT-20, HT 762.T, UACC-812, HCC1954 Hs 574.T BT-483 BT-549, DU4475, Hs 578T, BT-474, HCC1806, UACC-893, HCC38, HCC70, HCC202, HCC1143, HCC1187, HCC1395, HCC1419, HCC1500, HCC1599, HCC1937, HCC2157, HCC2218, HCC1569
Ovarian Cancer: Caov-3, TOV-21G, Hs 38.T, Hs 571.T, ES-2, TE 84.T
Pancreatic Cancer: BxPC-3, HPAF-II, HPAC, Panc 03.27, Panc 08.13, Panc 02.03, Panc 02.13, Panc 04.03, Panc 05.04, Cagan-2, CFPAC-1, PL45, Panc 10.05, MIA PaCa-2, PANC-1
Lung Cancer: Hs 229.T, NCI-H2135, NCI-H2172, NCI-H2444, NCI-H835, UMC-11, NCI-H727, NCI-H720, Hs 573.T, NCI-H596 NCI-H1688, NCI-H1417, NCI-H1836, NCI-H1672 HLF-a, NCI-H292, NCI-H2126, Calu-6, NCI-H2170, NCI-H520, SW 900, Hs 57.T
Colorectal cancer: NCI-H716, NCI-H747, NCI-H508, NCI-H498, SNU-C2B, SNU-C2A, LS513, LS1034, LS411N, WiDr, COLO 320DM, COLO 320HSR, DLD-1, HCT-15, SW480, SW403, SW48, SW1116, SW948, SW1417, LS123, LS 180, LS 174T, C2BBe1, Hs 257.T, Hs 587.Int, Caco-2, HT-29, HCT 116, ATRFLOX, SW1463, Hs 200.T, Hs 219.T, Hs 722.T.
Non-small cell lung cancer: NCI-H1581 NCI-H23, NCI-H522, NCI-H1435, NCI-H1563, NCI-H1651, NCI-H1734, NCI-H1793, NCI-H1838, NCI-H1975, NCI-H2073, NCI-H2085, NCI-H2228, NCI-H2342, NCI-H2347, NCI-H2066, NCI-H2286, NCI-H1703, SW 1573, NCI-H358, NCI-H810, DMS 79, DMS 53, DMS 114, SW 1271, NCI-H2227, NCI-H1963, SHP-77, H69AR
Skin Cancer: 182-PF, SK 166-ME, SK, TE 354.T, A-431, A431NS, A253*, Hs 357.T, Hs 941.T, Hs 295.T, Hs 63.T, Hs 892.T, Hs 898.T, Hs 416.T, Hs 925.T, Hs 156.T, WM-115, Hs 600.T, Hs 688(A).T, Hs 839.T, Hs 852.T, Hs 906(A).T, Hs 906(B).T, Hs 908.Sk, Hs 936.T, Hs 936.T (C1), Hs 939.T, A101D CHL-1, HMCB (Human Melanoma Cell Bowles), C32TG, C32, G-361, A-375, A375.S2, COLO 829, Hs 940.T, HT-144, Malme-3M, RPMI-7951, SK-MEL-5, SK-MEL-24, SK-MEL-28 SK-MEL-31, WM278,451Lu, WM1552C, WM35, WM793B, 1205Lu, WM39, A7
Liver Cancer: C3A, SNU-398, SNU-449, SNU-182, SNU-475, Hep 3B2.1-7, Hep G2, SNU-387, SNU-423, PLC/PRF/5
Brain cancer: A172, U-138 MG, DBTRG-05MG, LN-18, LN-229, U-87 MG, U-118 MG, M059K, M059J, LNZTA3WT4, LNZTA3WT11, Hs 683, PFSK-1, CHP-212, IMR-32, H4 Bone/Bone Marrow cancer: Hs 819.T, SW 1353, TF-1, TF-1a, TF-1.CN5a.1, HEL 92.1.7, KG 1, Hs 709.T, Hs 454.T, NCI-H929, 143.98.2, G-292, done A141B1, MG-63, HOS, KHOS/NP (R-970-5), KHOS-240S, KHOS-321H, MNNG/HOS (CI #5), Hs 3.T, Hs 39.T, Hs 184.T, Hs 188.T, Hs 387.T, Hs 704.T, Hs 707(A).T, Hs 735.T, Hs 755(B).T, Hs 781.T, Hs 792(B).T, Hs 805.T, Hs 811.T, Hs 866.T Hs 870.T, Hs 871.T, Hs 889.T, Hs 890.T, R-970-5, TE 417.T, TE 418.T, TO 203.T, HT 728.T, Hs 14.T, T1-73, 143B, 143B PML BK TK, Saos-2, U-2 OS, Hs 88.T, Hs 864.T, SJSA-1, Hs 900.T, Hs 903.T, Hs 919.T, SK-ES-1, Hs 706.T, Hs 737.T, Hs 821.T, Hs 846.T, Hs 883.T Hs 822.T, Hs 863.T, RD-ES, TE 76.T, TE 130.T, Hs 814.T, Hs 324.T, SW 982, MEG-01
Blood cancer: SUP-B15, CCRF-SB, 8E5, TALL-104, MOLT-4, CCRF-CEM, CCRF-HSB-2, MOLT-3, CEM/C2, CEM/C1, THP-1 TIB-202, AML-193, Kasumi-1 Kasumi-3, BDCM, AML14.3D10/CCCKR3 Clone 16, Kasumi-6, HL-60, Clone 15 HL-60, HL-60/MX2, HL-60/MX1, J.CaM1.6, Jurkat, Clone E6-1, J.RT3-T3.5, D1.1, J45.01, MV-4-11, Kasumi-4, KU812, KU812E, KU812F, RPMI 6666, U266B1, RPMI 8226, Mo, Mo-B, SUP-T1, JM1, GDM-1, CESS, ARH-77,1A2, H9/HTLV-IIIB, HuT 78, JSC-1, BCP-1,2B8, Daudi, EB-3, Raji, Jiyoye, NAMALWA, HS-Sultan, CA46, GA-10, GA-10 (Clone 4), GA-10 (Clone 20), NC-37,20B8, HKB-11,1G2, HH, H9, MJ, BC-1, BC-2, Toledo, U-937, TUR, DB, BC-3
Sarcoma: TE 441.T, TE 617.T, Hs 729.T, TE 381.T, RD, A-673, Hs 729, A-204, Hs 94.T, Hs 132.T, Hs 127.T, Hs 701.T, HT-1080, Hs 778(A).T, Hs 778(B).T, Hs 15.T SW 684, TE 115.T, Hs 93.T, Hs 934.T, Hs 935.T
Lymph node Cancer: Hs 604.T, Hs 751.T, Hs 445, Hs 611.T, Hs 616, Hs 505.T, Hs 491.T